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RIUNNE TD Duarte ER.Pdf (2.970Mb) “APLICACIÓN DE HERRAMIENTAS BIOTECNOLÓGICAS EN LA MICROPROPAGACIÓN DE ESPECIES ARBÓREAS NATIVAS DE LA SELVA MISIONERA EN PELIGRO DE EXTINCIÓN” EVELYN RAQUEL DUARTE Tesis para optar el grado de Doctor de la Universidad Nacional del Nordeste en el Área de Recursos Naturales UNIVERSIDAD NACIONAL DEL NORDESTE Facultad de Ciencias Agrarias 2015 RESUMEN La Selva Misionera es considerada una de las eco-regiones del país con mayor diversidad biológica. Sin embargo, la explotación extractiva de sus principales especies arbóreas maderables y la falta de acciones complementarias de manejo, provocan un alto grado de erosión genética que compromete la sustentabilidad del sistema; requiriéndose de acciones concretas que promuevan la restauración de la biodiversidad mediante la implantación de las especies de mayor consumo. En este contexto, el objetivo general del trabajo fue desarrollar procedimientos de propagación masiva que permitan la obtención de plantas de las especies amenazadas o en peligro de extinción. A partir de la recolección de frutos y semillas de ejemplares de Balfourodendron riedelianum (guatambú blanco), Cedrela fissilis (cedro misionero), Cordia trichotoma (peteribí), Handroanthus heptaphyllus (lapacho rosado) y Myrocarpus frondosus (incienso) que crecen en la Reserva de Usos Múltiples de la Facultad de Ciencias Forestales (Universidad Nacional de Misiones), se desarrollaron sendos experimentos que permitieron el establecimiento in vitro y germinación de las semillas de la mayoría de las especies estudiadas, excepto M. frondosus; las que fueron utilizadas como fuente dadora de explantes en los ensayos de organogénesis y micropropagación. Los resultados obtenidos indican la posibilidad de propagar estas especies ya sea a través de un proceso de organogénesis indirecta (B. riedelianum), micropropagación a partir del cultivo in vitro de segmentos caulinares (C. fissilis, C. trichotoma, H. heptaphyllus); o bien, por combinación de ambas técnicas (B. riedelianum), cuyas plantas resultantes se adaptan rápidamente a las condiciones ambientales externas durante la fase de aclimatación. ABSTRACT The Misiones rainforest is considered one of the eco-regions with the greatest biodiversity. However, the extractive exploitation of its main timber tree species and the lack of additional management actions, cause a high degree of genetic erosion that compromises the system sustainability; requiring specific actions to promote restoration of biodiversity by implanting the most widely consumed species. Along these lines, the main objective was developing procedures of mass propagation that will produce plants from endangered species. Fruits and seeds were collected from Balfourodendron riedelianum Cedrela fissilis, Cordia trichotoma, Handroanthus heptaphyllus, and Myrocarpus frondosus wild plants growing in the wildlife reserve (Facultad de Ciencias Forestales, Universidad Nacional de Misiones). The obtained results allowed the in vitro establishment and germination of seed from the majority of the studied species, except M. frondosus, which were used as source of explants in the assays of organogenesis and micropropagation. The results indicate the possibility of spreading these species either through a process of indirect organogenesis (B. riedelianum), micropropagation from in vitro culture of stem-segments (C. fissilis, C. trichotoma, H. heptaphyllus); or, by combining both techniques (B. riedelianum). The propagated plants swiftly adapt to the external environmental conditions during acclimation. Los experimentos llevados a cabo en esta tesis han sido realizados en el Instituto de Botánica del Nordeste (IBONE-CONICET), Facultad de Ciencias Agrarias (UNNE). DIRECTOR DE TESIS: Dr. Pedro A. Sansberro CO-DIRECTORA DE TESIS: Dra. Claudia V. Luna Queda prohibido no sonreír a los problemas, no luchar por lo que quieres, abandonarlo todo por miedo y no convertir en realidad tus sueños. Pablo Neruda Dedicado a A Mi madre por ser mi apoyo y fortaleza incondicional en toda mi vida A mis sobrinos Kiara, Agustina, Ulises, Gabriel, Brhiana, Emily y Santino quienes con su cariño me brindaron la fuerza que necesitaba para seguir adelante. AGRADECIMIENTOS A Dios por estar a mi lado y darme la fuerza y sabiduría que me permitió concluir con esta etapa de mi vida, ya que sin su bendición no habría sido posible. A mi madre, quien ocupando el lugar de padre y madre me ha dado el empuje y ha sido siempre mi soporte incondicional frente a cualquier problema que se presentara. Por ser un ejemplo de mujer, manteniéndose siempre firme ante sus convicciones, olvidando muchas veces su propia felicidad. A mis hermanos Adrián, Javier, Hugo y Daniel quienes me ayudaron y acompañaron siempre. A mi Director de tesis el Dr. Pedro Sansberro quien confió en mí, y dedicó tanto tiempo de su vida enseñándome y guiándome en camino, y hacer todo lo posible para concluir esta tesis. A mi Co-Directora la Dra. Claudia Luna que además de enseñarme y guiarme en la realización de esta tesis, también me brindó cariño y una gran amistad, convirtiéndose en un apoyo fundamental para poder lograr esta tesis. Al Dr. Luis Mroginski, por darme la oportunidad de ejecutar este proyecto en el Instituto de Botánica del Nordeste para lo cual brindó el equipamiento e infraestructura del Laboratorio de Fisiología Vegetal a su cargo. Al Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) por la beca otorgada para realizar esta tesis. A los profesionales y técnicos de la Facultad de Ciencias Agrarias e Instituto de Botánica del Nordeste por la colaboración prestada en todo momento. A la Universidad Nacional del Nordeste. A la Facultad de Ciencias Agrarias. Al Banco Regional de Semillas de la Facultad de Ciencias Forestales por ceder semillas de guatambú, lapacho e incienso. A la Dra. Ana María Gonzalez por su asesoramiento durante realización de los cortes anatómicos. A la Dra. Carolina Peichoto por su asesoramiento en cuestiones botánicas. A Dra. Ernestina Galdeano por su asesoramiento y apoyo en la identificación del insecto Amblycerus longesuturalis. A la Ing. Ftal. Beatriz Eibl por su asesoramiento en cuestiones fisiológicas de semillas. A la Dra. Adriana Salinas y al Dr. Collante por realizar imágenes radiográficas en las semillas de Cordia trichotoma. A la Familia Mattes quienes cedieron gentilmente la posibilidad de cosechar semillas de los árboles semilleros de su propiedad. A mis compañeros de laboratorio Maximiliano, Fabiana, Laura, Natalia, Edgardo, Gabriela, Candela, María Laura, Laurita, María Antonia, Francisca, Ricardo, Mónica, Eduardo F., Graciela, Eduardo M., Patricia y Mirta por el apoyo brindado durante la realización de este proyecto. A mis amigos Claudio, Guillermo, Diego, Daily, Liliana, Pablo, Luis y César. A todas aquellas personas que de una u otra forma colaboraron en la concreción de esta tesis. A todos ustedes que caminaron junto a mí en este periodo de mi vida y me brindaron su valioso tiempo, conocimiento y cariño. ¡MUCHAS GRACIAS! PRODUCCIÓN CIENTÍFICA Trabajo Publicado Detección de daño del coleóptero Amblycerus longesuturalis para la selección y germinación in vitro de semillas de Peteribí (Cordia trichotoma [Vell.] Arrab. ex Steudel). Duarte, E.; Acevedo, M; Sansberro, P; Luna, C. Revista de la Facultad de Agronomía de La Plata 112 (1): 18-27. 2014. Trabajos Enviados para su Publicación In vitro propagation of Handroanthus heptaphyllus (Vell.) Mattos by shoot cultures. Por: Duarte, E; Sansberro, P; Luna C. Revista: Trees - Structure and Function. Efecto de la testa sobre la germinación de semillas de Handroanthus heptaphyllus tras distintos tiempos de almacenamiento. Por: Duarte, E; Avico, E; Sansberro, P; Luna, C. Revista Ciencias Agronómicas. Universidad Nacional de Rosario. Presentaciones a Congresos Internacionales Direct shoot regeneration from mature zygotic embryos of Pinus elliottii x P. caribaea hybrids and plantlets production assisted by temporary immersion bioreactors. Por: Fortes, N; Duarte, E; González, A; Sansberro, P; Luna, C. 2014. IUFRO Working Party 2.09.02 - Somatic Embryogenesis and Other Vegetative Propagation Technologies. Vitoria-Gasteiz, España. Presentaciones a Congresos Nacionales y Regionales Uso de Biorreactores de inmersión temporal (BIT) para promover la rizogénesis adventicia de una especie arbórea Duarte, E; Sansberro, P; Luna, C. 2013.VIII Simposio Nacional de Biotecnología REDBIO Argentina. Regeneración in vitro de yemas adventicias a partir del cultivo de segmentos caulinares de Cedrela fissilis. Duarte, E; Luna, C; Sansberro, P. 2011.VII Simposio Nacional de Biotecnología REDBIO Argentina. Presentaciones en Reuniones Científicas Métodos comparativos de determinación de Viabilidad en Semillas Forestales Nativas. Barone, J; Duarte, E; Luna, C. 2014. XXIII Reunión de Comunicaciones Científicas, Técnicas y de Extensión (FCA-UNNE). Germinación in vitro de Handroanthus heptaphyllus (Vell.) Mattos (Bignoniaceae). Duarte, E; Luna, C; Sansberro, P. 2011. XXII Reunión de Comunicaciones Científicas, Técnicas y de Extensión (FCA-UNNE). Morfometría de semillas de Cordia Trichotoma (Vell.) Arrab. Ex. Steudel de dos diferentes procedencias. López, E; Duarte, E; Luna, C. 2014. XX Reunión de Comunicaciones Científicas, Tecnológicas (SGCyT –UNNE). XIX Reunión de Comunicaciones Científicas, Tecnológicas (SGCyT –UNNE). Germinación in vitro de Handroanthus heptaphyllus (Vell.) Mattos (Bignoniaceae). Duarte, E; Luna, C; Sansberro, P. 2012. Germinación in vitro de Handroanthus heptaphyllus (Vell.) Mattos (Bignoniaceae). Duarte,
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